SIRT 1通过下调PPAR-γ受体提升人乳牙牙髓干细胞骨向分化效能  

SIRT1 Enhances Osteogenesis in Stem Cells from Human Exfoliated Deciduous Teeth via Downregulation of Peroxisome Proliferator-Activated Receptor

作　　者：曹威[1] 刘梦佳 张清彬[1] CAO Wei;LIU Mengjia;ZHANG Qingbin(Department of Temporomandibular Joint, Affiliated Stomatology Hospital of Guangzhou Medical University, Guangdong Engineering Research Center of Oral Restoration and Reconstruction, Guangzhou Key Laboratory of Basic and Applied Research of Oral Regenerative Medicine, Guangzhou 510182, China;Department of Anatomy, Guangzhou Medical University, Guangzhou 511436, China.)

机构地区：[1]广州医科大学附属口腔医院颞下颌关节科·广东省口腔组织修复与重建工程技术研究中心·广州市口腔再生医学基础与应用研究重点实验室,广东广州510182 [2]广州医科大学基础医学院人体解剖学教研室,广东广州511436

出　　处：《口腔医学研究》2022年第2期169-175,共7页Journal of Oral Science Research

基　　金：广州市卫生健康科技一般引导项目(编号:20191A010067);广州市卫生健康委特色技术项目(编号:2019TS42)。

摘　　要：目的:研究SIRT1对人乳牙牙髓干细胞(SHEDs)成骨分化效能的影响。方法:选取特异性SIRT1激活剂SRT1720和抑制剂EX527,预处理SHEDs 3 d后作为工作细胞用于实验。CCK-8实验检测SRT1720和EX527与SHEDs共培养1、2、3、7 d后的细胞增殖能力变化。筛选出最佳浓度为0.5μmol/L的SRT1720和EX527溶液,利用成骨诱导培养基诱导SHEDs骨向分化。通过碱性磷酸酶染色及定量检测,茜素红矿化结节染色等方法分别检测早、晚期成骨分化,通过RT-PCR检测骨向分化标志物ALP、Runx2、骨桥蛋白、Ⅰ型胶原、骨钙蛋白以及SIRT1、PPAR-γ等相关基因及蛋白表达水平,研究SIRT1调控SHEDs体外骨向分化的可能机制。结果:SIRT1激活剂组骨向分化标志物基因mRNA较SIRT1抑制剂组增强,表明随着SIRT1表达上调,SHEDs的成骨分化效能也一并上调,而PPAR-γ受体相应下调。结论:SIRT1的表达与SHEDs的骨向分化能力呈正相关,SIRT1促进骨向分化的作用与BMP信号通路中的PPAR-γ受体下调密切相关。Objective:To identify the role of SIRT1 on osteogenic differentiation of stem cells from human exfoliated deciduous teeth.Methods:A defined concentration of SIRT1 activator(SRT1720)and inhibitor(EX527)is 0.5μmol/L.Then SRT1720 and EX527 were added to the experiment groups for preconditioning 3 days.We investigated the effects of SIRT1 on the osteogenic differentiation of SHEDs.Expression of osteoblastic differentiation markers was assessed by RT-PCR.Results:Activation of SIRT1 using SRT1720 increased the expression of osteoblastic differentiation markers,as assessed by alkaline phosphatase,Runt-related transcription factor-2,osteopontin,collagen type Iα1,and osteocalcin.In contrast,inhibition of SIRT1 using EX527 suppressed the expression of above-mentioned factors.Conclusion:SIRT1 is a key regulator of differentiation of stem cells from human exfoliated deciduous teeth.SIRT1 also regulated the expression of PPAR-γ.The SIRT1/PPAR-γpathway may be crucial for enhancement of osteogenesis.

关 键 词：SIRT1 人乳牙牙髓干细胞 PPAR-Γ 骨向分化 

分 类 号：R73[医药卫生—肿瘤]